Abstract
Diablo/Smac is a mammalian pro-apoptotic protein that can antagonize the inhibitor of apoptosis proteins (IAPs). We have produced monoclonal antibodies specific for Diablo and have used these to examine its tissue distribution and subcellular localization in healthy and apoptotic cells. Diablo could be detected in a wide range of mouse tissues including liver, kidney, lung, intestine, pancreas and testes by Western blot analysis. Immunohistochemical analysis found Diablo to be most abundant in the germinal cells of the testes, the parenchymal cells of the liver and the tubule cells of the kidney. In support of previous subcellular localization analysis, Diablo was present within the mitochondria of healthy cells, but released into the cytosol following the induction of apoptosis by UV.
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Abbreviations
- mAbs:
-
monoclonal antibodies
- IAPs:
-
inhibitor of apoptosis proteins
- BIR:
-
baculoviral IAP repeat
- s.c:
-
subcutaneous
- i.p:
-
intraperitonealy
- i.v:
-
intravenously
- FITC:
-
fluorescein-isothiocyanate
- HA:
-
epitope (YPYDVPDYA) tag
- HA Diablo:
-
epitope HA-tagged Diablo protein
- IP:
-
immunoprecipitation
- IF:
-
immunofluorescence
- WB:
-
Western blotting
- FACS:
-
fluorescence-activated cell sorter
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Acknowledgements
We would like to acknowledge L Cullen and K Davern for assistance with antibody production, V Lapatis for assistance with confocal microscopy, Dr. D Huang for providing valuable reagents and Dr(s). A Strasser, J Silke and PG Ekert for valuable technical advice. This work was supported by an NHMRC grant to the WEHI (Reg Key 973002) and by the Anti-Cancer Council of Victoria. DL Vaux is a scholar of the Leukemia and Lymphoma Society.
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Tikoo, A., O'Reilly, L., Day, C. et al. Tissue distribution of Diablo/Smac revealed by monoclonal antibodies. Cell Death Differ 9, 710–716 (2002). https://doi.org/10.1038/sj.cdd.4401031
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DOI: https://doi.org/10.1038/sj.cdd.4401031
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